title: Biochemical analysis of the role of G118R-linked dolutegravir drug resistance substitutions in HIV-1 integrase.
abstract: Drug resistance mutations (DRMs) have been reported for all currently approved anti-HIV drugs, including the latest integrase strand transfer inhibitors (INSTIs). We previously used the new INSTI dolutegravir (DTG) to select a G118R integrase resistance substitution in tissue culture and also showed that secondary substitutions emerged at positions H51Y and E138K. Now, we have characterized the impact of the G118R substitution, alone or in combination with either H51Y or E138K, on 3' processing and integrase strand transfer activity. The results show that G118R primarily impacted the strand transfer step of integration by diminishing the ability of integrase-long terminal repeat (LTR) complexes to bind target DNA. The addition of H51Y and E138K to G118R partially restored strand transfer activity by modulating the formation of integrase-LTR complexes through increasing LTR DNA affinity and total DNA binding, respectively. This unique mechanism, in which one function of HIV integrase partially compensates for the defect in another function, has not been previously reported. The G118R substitution resulted in low-level resistance to DTG, raltegravir (RAL), and elvitegravir (EVG). The addition of either of H51Y or E138K to G118R did not enhance resistance to DTG, RAL, or EVG. Homology modeling provided insight into the mechanism of resistance conferred by G118R as well as the effects of H51Y or E138K on enzyme activity. The G118R substitution therefore represents a potential avenue for resistance to DTG, similar to that previously described for the R263K substitution. For both pathways, secondary substitutions can lead to either diminished integrase activity and/or increased INSTI susceptibility.
authors.affiliation: McGill University AIDS Centre, Lady Davis for Medical Research, Jewish General Hospital, Montreal, Quebec, Canada.
authors.forename: Peter K
authors.initials: PK
authors.lastname: Quashie
authors.forename: Thibault
authors.initials: T
authors.lastname: Mesplède
authors.forename: Ying-Shan
authors.initials: YS
authors.lastname: Han
authors.forename: Tamar
authors.initials: T
authors.lastname: Veres
authors.forename: Nathan
authors.initials: N
authors.lastname: Osman
authors.forename: Said
authors.initials: S
authors.lastname: Hassounah
authors.forename: Richard D
authors.initials: RD
authors.lastname: Sloan
authors.forename: Hong-Tao
authors.initials: HT
authors.lastname: Xu
authors.forename: Mark A
authors.initials: MA
authors.lastname: Wainberg
mesh_terms: D024882:Drug Resistance, Viral / Q000235:genetics; D004789:Enzyme Activation / Q000187:drug effects; D019427:HIV Integrase / Q000235:genetics* / Q000378:metabolism; D019428:HIV Integrase Inhibitors / Q000494:pharmacology*; D006575:Heterocyclic Compounds, 3-Ring / Q000494:pharmacology*; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D010078:Oxazines; D010879:Piperazines; D011728:Pyridones; D000099226:Dolutegravir
chemical_list: D019427:HIV Integrase; D019428:HIV Integrase Inhibitors; D006575:Heterocyclic Compounds, 3-Ring; D010078:Oxazines; D010879:Piperazines; D011728:Pyridones; D000099226:Dolutegravir; C514801:p31 integrase protein, Human immunodeficiency virus 1